Project Abstract and Summary Alzheimer?s disease (AD) is a progressive neurodegenerative disease and is considered the predominate cause of dementia worldwide. In the United States (US), it is the sixth leading cause of death. It results in a loss of memory, thinking skills and task performance. The pathological hallmarks of AD are accumulated amyloid ? (A?) plaques and neurofibrillary tangles (NFTs). Studies have shown that reduction in wild-type tau prevents A?-dependent behavioral and cognitive deficits, suggesting that therapeutic interventions that alter the levels of tau may be beneficial. Our lab provided convincing evidence that either silencing the Sp1 gene, using small interfering RNA, or treatment of animals with Tolfenamic Acid (TA), lowers the expression of AD-related Sp1 target genes. TA (Clotam Rapid) is currently used in Europe and other countries to treat symptoms of migraine headaches. TA serves as an excellent candidate for the design and synthesis of novel derivatives. Thus, we synthesized a series of analogs using the lead compound scaffold by focusing on optimizing potency, selectivity, and generating drug-like molecules for CNS delivery, while, also, honing the physical properties of the compounds. Furthermore, we aimed to decrease the COX-related action of these compounds and enhance their targeting of Sp1/tau. We, now, have a series of compounds and we would like to examine: efficacy, potency, cytotoxicity, potential mechanism of action, selectivity and off target effects in an in vitro cell culture model. The following are the aims of the proposal: 1) Investigate the ability of analogs, with a better safety profile than TA, to lower the mRNA and protein levels of total tau, site specific tau phosphorylation, CDK5, Sp1, and COX2, in an in vitro model, 2) Testing of lead analogs for mode of action and target engagement, 3) Analyze the brain transfer potential of TA analogs using in silico and in vitro approaches. The expected outcomes are that we will find that TA analogs specifically target Sp1 and other related biomarkers by chelating Zn. We, also, expect that the analogs will cross the blood brain barrier (BBB) due to structural and physiochemical characteristics that enhance brain penetration potential.